Wong Alan, Li Xiaonan, Molin Laurent, Solari Florence, Elena-Herrmann Bénédicte, Sakellariou Dimitris
CEA Saclay, DSM, IRAMIS, UMR CEA/CNRS 3299-NIMBE, Laboratoire Structure et Dynamique par Résonance Magnétique, F-91191, Gif-sur-Yvette Cedex, France.
Anal Chem. 2014 Jun 17;86(12):6064-70. doi: 10.1021/ac501208z. Epub 2014 Jun 4.
Analysis of model organisms, such as the submillimeter-size Caenorhabditis elegans, plays a central role in understanding biological functions across species and in characterizing phenotypes associated with genetic mutations. In recent years, metabolic phenotyping studies of C. elegans based on (1)H high-resolution magic-angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy have relied on the observation of large populations of nematodes, requiring labor-intensive sample preparation that considerably limits high-throughput characterization of C. elegans. In this work, we open new platforms for metabolic phenotyping of C. elegans mutants. We determine rich metabolic profiles (31 metabolites identified) from samples of 12 individuals using a (1)H NMR microprobe featuring high-resolution magic-angle coil spinning (HR-MACS), a simple conversion of a standard HR-MAS probe to μHR-MAS. In addition, we characterize the metabolic variations between two different strains of C. elegans (wild-type vs slcf-1 mutant). We also acquire a NMR spectrum of a single C. elegans worm at 23.5 T. This study represents the first example of a metabolomic investigation carried out on a small number of submillimeter-size organisms, demonstrating the potential of NMR microtechnologies for metabolomics screening of small model organisms.
对诸如亚毫米大小的秀丽隐杆线虫等模式生物的分析，在理解跨物种生物功能以及表征与基因突变相关的表型方面发挥着核心作用。近年来，基于氢-高分辨率魔角旋转（HR-MAS）核磁共振（NMR）光谱的秀丽隐杆线虫代谢表型研究依赖于对线虫大量群体的观察，需要耗费大量人力的样品制备，这极大地限制了秀丽隐杆线虫的高通量表征。在这项工作中，我们为秀丽隐杆线虫突变体的代谢表型分析开辟了新平台。我们使用具有高分辨率魔角线圈旋转（HR-MACS）的氢NMR微探头，从12个个体的样本中确定了丰富的代谢谱（鉴定出31种代谢物），这是将标准HR-MAS探头简单转换为μHR-MAS探头。此外，我们表征了秀丽隐杆线虫两种不同菌株（野生型与slcf-1突变体）之间的代谢差异。我们还在23.5 T下获取了单个秀丽隐杆线虫的NMR光谱。这项研究代表了对少量亚毫米大小生物进行代谢组学研究的首个实例，证明了NMR微技术在小型模式生物代谢组学筛选中的潜力。
